title: Protein microarray characterization of the S-nitrosoproteome.
abstract: Nitric oxide (NO) mediates a substantial part of its physiologic functions via S-nitrosylation, however the cellular substrates for NO-mediated S-nitrosylation are largely unknown. Here we describe the S-nitrosoproteome using a high-density protein microarray chip containing 16,368 unique human proteins. We identified 834 potentially S-nitrosylated human proteins. Using a unique and highly specific labeling and affinity capture of S-nitrosylated proteins, 138 cysteine residues on 131 peptides in 95 proteins were determined, defining critical sites of NO's actions. Of these cysteine residues 113 are novel sites of S-nitrosylation. A consensus sequence motif from these 834 proteins for S-nitrosylation was identified, suggesting that the residues flanking the S-nitrosylated cysteine are likely to be the critical determinant of whether the cysteine is S-nitrosylated. We identify eight ubiquitin E3 ligases, RNF10, RNF11, RNF41, RNF141, RNF181, RNF208, WWP2, and UBE3A, whose activities are modulated by S-nitrosylation, providing a unique regulatory mechanism of the ubiquitin proteasome system. These results define a new and extensive set of proteins that are susceptible to NO regulation via S-nitrosylation. Similar approaches could be used to identify other post-translational modification proteomes.
authors.affiliation: Neuroregeneration Program, Institute for Cell Engineering.
authors.forename: Yun-Il
authors.initials: YI
authors.lastname: Lee
authors.forename: Daniel
authors.initials: D
authors.lastname: Giovinazzo
authors.forename: Ho Chul
authors.initials: HC
authors.lastname: Kang
authors.forename: Yunjong
authors.initials: Y
authors.lastname: Lee
authors.forename: Jun Seop
authors.initials: JS
authors.lastname: Jeong
authors.forename: Paschalis-Thomas
authors.initials: PT
authors.lastname: Doulias
authors.forename: Zhi
authors.initials: Z
authors.lastname: Xie
authors.forename: Jianfei
authors.initials: J
authors.lastname: Hu
authors.forename: Mehdi
authors.initials: M
authors.lastname: Ghasemi
authors.forename: Harry
authors.initials: H
authors.lastname: Ischiropoulos
authors.forename: Jiang
authors.initials: J
authors.lastname: Qian
authors.forename: Heng
authors.initials: H
authors.lastname: Zhu
authors.forename: Seth
authors.initials: S
authors.lastname: Blackshaw
authors.forename: Valina L
authors.initials: VL
authors.lastname: Dawson
authors.forename: Ted M
authors.initials: TM
authors.lastname: Dawson
mesh_terms: D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism*; D040081:Protein Array Analysis; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011506:Proteins / Q000378:metabolism; D020543:Proteome
chemical_list: D011506:Proteins; D020543:Proteome; D009569:Nitric Oxide